Preparation And Storage
Recommended Assay Procedures
7A6 may be used for western blot analysis of NF-ATc1 (1-2 µg/ml). Note that while the predicted M W of NF-ATc is ~100 kDa, the observed M W may range between 100 to 120 kDa due to post-translational modification of NF-ATc. Other applications include immunoprecipitation (2 µg/ml), mobility shift assays (EMSA) and immunofluorescence microscopy of paraformaldehyde-fixed, tissue cultured cells. Jurkat human T cells (ATCC TIB-152) are suggested as a positive control for expression of NF-ATc.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Companion Products
The NF-AT family of transcription factors are regulators of early immune response genes in T cells following their activation by CD40L, FasL and nterleukins including IL-2, -3, -4 and -5. NF-AT proteins function in both signal transduction and transcription control. The DNA binding, active NF-AT complex contains a cytoplasmic, Ca2+/calcineurin dependent, cyclosporin sensitive subunit, designated NF-ATc, which is present in the cytoplasm of resting cells. Upon cell activation, NF-ATc is dephosphorylated and thus activated by calcineurin, resulting in rapid translocation of NF-ATc to the nucleus. The active NF-AT complex also contains a nuclear component, NF-ATn. In vitro, NF-AT cooperates with the mitogenic transcription factor AP-1 to induce multiple cytokine genes. NF-ATc is encoded by four genes: NF-ATc1 (originally named NF-ATc), NF-ATc2 (originally named NF-ATp), NF-ATc3 (NF-AT3) and NF- ATc4 (NF-AT4). These proteins are differentially expressed in tissues, suggesting that each may activate distinct sets of genes. NF-ATc1 is expressed in lymphoid organs and is upregulated in activated T cell and thymocytes. NF-ATc1 is required for the development of T cells and has been shown to contribute to the production of IL-2 and the development of Th2 responses.
7A6 recognizes human and mouse NF-ATc1. The antibody recognizes a region between amino acids 197 to 304 of human NF-ATc1, which lies outside the region of homology between NF-ATc1 and NF-ATc2.3 Please note that in the original publication for the antibody (Northrop et al. 1994), the clone name 7A6 is not specifically identified.
Development References (5)
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Crabtree GR. Contingent genetic regulatory events in T lymphocyte activation. Science. 1989; 243(4889):355-361. (Biology). View Reference
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Northrop JP, Ho SN, Chen L. NF-AT components define a family of transcription factors targeted in T-cell activation. Nature. 1994; 369(6480):497-502. (Clone-specific: Immunoprecipitation). View Reference
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Timmerman LA, Clipstone NA, Ho SN, Northrop JP, Crabtree GR. Rapid shuttling of NF-AT in discrimination of Ca2+ signals and immunosuppression. Nature. 1996; 383(6603):837-840. (Clone-specific: Immunofluorescence, Immunoprecipitation). View Reference
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Timmerman LA, Healy JI, Ho SN, Chen L, Goodnow CC, Crabtree GR. Redundant expression but selective utilization of nuclear factor of activated T cells family members. J Immunol. 1997; 159(6):2735-2740. (Clone-specific: Immunoprecipitation). View Reference
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Yoshida H, Nishina H, Takimoto H. The transcription factor NF-ATc1 regulates lymphocyte proliferation and Th2 cytokine production. Immunity. 1998; 8(1):115-124. (Biology). View Reference
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